Materials and methods for improving lung function and for prevention and/or treatment of radiation-induced lung complications

ABSTRACT

The subject invention provides therapeutic compositions and uses thereof for improving pulmonary function. In one embodiment, the therapeutic composition comprises one or more free amino acids selected from lysine, glycine, threonine, valine, tyrosine, aspartic acid, isoleucine, tryptophan, asparagine, and serine; and electrolytes. In one embodiment, the subject invention can be used to prevent or treat long-term lung complications induced by radiation.

CROSS-REFERENCE TO A RELATED APPLICATION

This application is a Continuation Application of U.S. application Ser.No. 16/508,854, filed Jul. 11, 2019, which is a Continuation Applicationof U.S. application Ser. No. 14/773,960, now U.S. Pat. No. 10,350,185,filed Sep. 9, 2015, which is a U.S. National Stage Application ofInternational Application No. PCT/US2014/023363, filed Mar. 11, 2014,which claims the benefit of U.S. provisional application Ser. No.61/775,754, filed Mar. 11, 2013, all of which are incorporated herein byreference in their entirety.

BACKGROUND OF INVENTION

Radiation therapy, a common treatment for malignancies, can cause severedamage to the lung—a highly radiosensitive organ. Radiation can cause abroad spectrum of pneumopathies, including acute-phasealveolitis/pneumonitis, late-stage chronic pulmonary fibrosis, andvarious respiratory dysfunctions such as dyspnea and pulmonary edema.

During the acute phase of radiation-induced lung injury, inflammation isthe predominant histological and physiologic feature. The initial injuryto lung tissues results in infiltration of inflammatory cells, such asmacrophages and neutrophils; focal accumulation of mononuclear cells;increased levels of inflammatory cytokines such as transforming growthfactor-β (TGF-β), interleukin-1α (IL-1α), and tumor necrosis factor(TNFα); and a decline in pulmonary function.

Radiation also induces late-stage pulmonary fibrosis—an insidiousfibroproliferative condition characterized by a gradual, irreversiblereplacement of normal parenchyma cells with fibrous, connective, matrixmacromolecules (e.g., collagens, fibronectins and proteoglycans) on andwithin the lungs, usually at sites of injury or infection. The excessiveformation of fibrous tissue, resulting from the activation andproliferation of fibroblast cells, destructs normal lung structure andfunction. For instance, the accumulation of fibrous tissue thickensalveolar walls, obliterates air space, and causes epithelial injury oreven alveolar collapse.

Patients suffering from pneumopathies (such as pneumonitis and pulmonaryfibrosis) experience a varying degree of exertional dyspnea, and in latestages, orthopnea, cyanosis, and respiratory failure. Currently, thereis no cure for radiation-induced pulmonary fibrosis. Median survival ofradiation-induced pulmonary fibrosis is about 2-3 years.

Radiation-induced pneumopathy not only causes devastating effects on thequality of patient life, but sometimes can be even more life-threateningthan the primary tumor or cancer. Consequently, the risk ofradiation-induced pneumopathy, such as pulmonary fibrosis, has become amajor dose-limiting factor and sometimes even prevents the use ofradiation therapy.

There is a need for therapeutic formulations for prevention andtreatment of radiation-induced lung injury and complications. As will beclear from the disclosures that follow, these and other benefits areprovided by the subject invention.

BRIEF SUMMARY

The subject invention provides materials and methods for improvingpulmonary function. In one embodiment, the subject invention is usefulfor the prevention and/or treatment of radiation-induced lung injury andlung complications, including radiation-induced alveolitis, pneumonitis,and pulmonary fibrosis.

In one embodiment, a composition of the subject invention is formulatedfor oral administration. In another embodiment, the composition isformulated for pulmonary administration.

In a preferred embodiment, the subject invention provides a method forimproving pulmonary function, and/or for the prevention and/or treatmentof radiation-induced lung injury and lung complications, wherein themethod comprises administering, to a patient or subject in need of suchtreatment, an effective amount of a composition comprising, consistingessentially of, or consisting of L-lysine, L-glycine, L-threonine,L-valine, L-tyrosine, L-aspartic acid, L-isoleucine, and L-serine; oneor more electrolytes selected from Na⁺, Ca²⁺, Mg²⁺, HCO₃ ⁻, and Cl⁻; andoptionally, therapeutically acceptable carriers, buffering agents, andflavoring agents.

In one embodiment, the total osmolarity of the therapeutic compositionis from about 165 mOsm to 300 mOsm, or any value therebetween. In oneembodiment, the composition has a pH from about 2.0 to 8.6, or any valuetherebetween.

In one embodiment, the subject invention can be used to prevent and/ortreat lung complications induced by radiation. In one specificembodiment, the subject invention can be used to prevent and/or treatlung complications induced by ionizing radiation. In certainembodiments, the subject invention can be used to prevent and/or treatradiation-induced lung complications including, but not limited to,alveolitis, pneumonitis, and pulmonary fibrosis.

In certain embodiments, the present invention can be used to treat lungdiseases including bronchial asthma, pneumonia, bronchiectasis,interstitial lung diseases, acute and/or chronic pneumonitis, chronicobstructive pulmonary disease (COPD), asthma, silicosis, and lunginjury.

BRIEF DESCRIPTION OF DRAWING

FIG. 1A-H shows the results of pulmonary function tests. Briefly, micereceived radiation at a dose of 8 Gy. Twenty-four hours afterirradiation, mice are treated with a therapeutic composition of thesubject invention for a period of 14 days. Six months after irradiation,mice treated in accordance with the subject invention have improvedfunction, when compared to control. The data show that the therapeuticcomposition improves pulmonary function and can be used to treatlong-term lung complications induced by radiation.

DETAILED DISCLOSURE

The subject invention provides therapeutic compositions and methods forimproving pulmonary function. In one embodiment, the subject inventionis useful for the prevention and/or treatment of radiation-induced lunginjury and lung complications, including radiation-induced alveolitis,pneumonitis, and pulmonary fibrosis.

In one embodiment, the composition is formulated for oraladministration. In another embodiment, the composition is formulated forpulmonary administration.

In a preferred embodiment, the subject invention provides a method forimproving pulmonary function, and/or for the prevention and/or treatmentof radiation-induced lung injury and lung complications, wherein themethod comprises administering, to a patient or subject in need of suchtreatment, an effective amount of a composition comprising, consistingessentially of, or consisting of L-lysine, L-glycine, L-threonine,L-valine, L-tyrosine, L-aspartic acid, L-isoleucine, and L-serine; oneor more electrolytes selected from Na⁺, Ca²⁺, Mg²⁺, HCO₃ ⁻, and Cl⁻; andoptionally, therapeutically acceptable carriers, buffering agents, andflavoring agents.

The composition can be administered to a patient or subject immediatelybefore, during, and/or after injury to the lungs, and can beadministered once or multiple times each day.

Advantageously, in one embodiment, the compositions of the subjectinvention can be used to prevent or treat radiation-induced long-termlung complications. In one embodiment, mice that received radiation at adose of 8 Gy are treated with the composition of the subject inventionstarting from 24 hours after irradiation, for a period of 14 days. Sixmonths after irradiation, pulmonary function test, electrophysiology,radiological and histopathological examinations are performed. Micetreated with a composition of the subject invention exhibit improvedpulmonary function, electrophysiology, radiological, andhistopathological features, when compared to control.

Therapeutic Compositions for Improving Pulmonary Function

In one embodiment, the subject invention provides a therapeuticcomposition for improving pulmonary function, wherein the compositioncomprises, consists essentially of, or consists of one or more freeamino acids selected from lysine, glycine, threonine, valine, tyrosine,aspartic acid, isoleucine, tryptophan, asparagine, and serine; andoptionally, therapeutically acceptable carriers, electrolytes, bufferingagents, and flavoring agents.

In one embodiment, the subject invention provides a therapeuticcomposition for improving pulmonary function, wherein the compositioncomprises, consists essentially of, or consists of lysine, glycine,threonine, valine, tyrosine, aspartic acid, isoleucine, tryptophan, andserine; and optionally, therapeutically acceptable carriers,electrolytes, buffering agents, and flavoring agents.

In another embodiment, the subject invention provides a therapeuticcomposition for improving pulmonary function, wherein the compositioncomprises, consists essentially of, or consists of lysine, glycine,threonine, valine, tyrosine, aspartic acid, isoleucine, and serine; andoptionally, therapeutically acceptable carriers, electrolytes, bufferingagents, and flavoring agents.

In one embodiment, the subject invention provides a therapeuticcomposition for improving pulmonary function, wherein the compositioncomprises, consists essentially of, or consists of one or more freeamino acids selected from L-lysine, L-glycine, L-threonine, L-valine,L-tyrosine, L-aspartic acid, L-isoleucine, L-tryptophan, L-asparagine,and L-serine; and optionally, therapeutically acceptable carriers,electrolytes, vitamins, buffering agents, and flavoring agents.

In one embodiment, the subject invention provides a therapeuticcomposition for improving pulmonary function, wherein the compositioncomprises, consists essentially of, or consists of L-lysine, L-glycine,L-threonine, L-valine, L-tyrosine, L-aspartic acid, L-isoleucine,L-tryptophan, and L-serine; and optionally, therapeutically acceptablecarriers, electrolytes, buffering agents, and flavoring agents.

In another embodiment, the subject invention provides a therapeuticcomposition for improving pulmonary function, wherein the compositioncomprises, consists essentially of, or consists of L-lysine, L-glycine,L-threonine, L-valine, L-tyrosine, L-aspartic acid, L-isoleucine, andL-serine; and optionally, therapeutically acceptable carriers,electrolytes, buffering agents, and flavoring agents.

In one embodiment, the free amino acids contained in the therapeuticcomposition can be present in neural or salt forms.

In one embodiment, the therapeutic composition further comprises one ormore electrolytes selected from Na⁺, Ca²⁺, Mg²⁺, HCO₃ ⁻, CO₃ ²⁻, andCl⁻.

In one embodiment, the total osmolarity of the composition is from about165 mOsm to 300 mOsm, or any value therebetween including, but notlimited to, 230 mOsm to 280 mOsm, 250 mOsm to 260 mOsm, and 200-220mOsm. In another embodiment, the composition has a total osmolarity thatis any value lower than 165 mOsm.

In certain embodiments, each free amino acid can be present at aconcentration from 4 mM to 40 mM, or any value therebetween, wherein thetotal osmolarity of the composition is from about 230 mOsm to 280 mOsm.Alternatively, if the amino acid concentration is calculated based onmg/l, each free amino acid can be present at a concentration from 100mg/l to 8000 mg/L, or any value therebetween, wherein the totalosmolarity of the composition is from about 240 mOsm to 280 mOsm.

In certain specific embodiments, the therapeutic composition comprisesone or more free amino acids present at their respective concentrationsas follows: lysine at a concentration of about 730 to 6575 mg/l, or anyvalue therebetween; aspartic acid at a concentration of about 532 to4792 mg/l, or any value therebetween; glycine at a concentration ofabout 300 to 2703 mg/l, or any value therebetween; isoleucine at aconcentration of about 525 to 4722 mg/l, or any value therebetween;threonine at a concentration of about 476 to 4288 mg/l, or any valuetherebetween; tyrosine at a concentration of about 725 to 6523 mg/l, orany value therebetween; valine at a concentration of about 469 to 4217mg/l, or any value therebetween; tryptophan at a concentration of about817 to 7352 mg/l, or any value therebetween; asparagine at aconcentration of about 528 to 4756 mg/l, or any value therebetween;and/or serine at a concentration of about 420 to 3784 mg/l, or any valuetherebetween; wherein the total osmolarity of the composition is fromabout 165 mOsm to 300 mOsm, or any value therebetween.

In certain specific embodiments, the therapeutic composition comprisesone or more free amino acids present at their respective concentrationsas follows: lysine at a concentration of about 730 to 6575 mg/l, or anyvalue therebetween; aspartic acid at a concentration of about 532 to4792 mg/l, or any value therebetween; glycine at a concentration ofabout 300 to 2703 mg/l, or any value therebetween; isoleucine at aconcentration of about 525 to 4722 mg/l, or any value therebetween;threonine at a concentration of about 100 to 4288 mg/l, or any valuetherebetween; tyrosine at a concentration of about 725 to 6523 mg/l, orany value therebetween; valine at a concentration of about 469 to 4217mg/l, or any value therebetween; and/or serine at a concentration ofabout 420 to 3784 mg/l, or any value therebetween; wherein the totalosmolarity of the composition is from about 165 mOsm to 300 mOsm, or anyvalue therebetween.

In one embodiment, the subject invention provides a formulationcomprising the following constituents: lysine (11-21 mOsm), asparticacid (3-13 mOsm), glycine (19-29 mOsm), isoleucine (19-29 mOsm),threonine (19-29 mOsm), tyrosine (0.5-5 mOsm), valine (19-29 mOsm),tryptophan (5-20 mOsm), asparagine (3-13 mOsm), and/or serine (3-8mOsm), or a subset of these ingredients.

In one embodiment, the composition has a pH from about 2.0 to 8.6, orany value therebetween. In certain embodiments, the composition has a pHfrom about 2.0 to 5.0, or any value therebetween, including, such as 2.0to 4.2 and 2.0 to 3.6. In certain embodiments, the composition has a pHfrom about 7.3 to 7.5, or any value therebetween including, such asabout 7.4. In certain embodiments, the composition has a pH from about4.0 to 8.5, or any value therebetween including, such as 5.0 to 8.2, 6.0to 8.0, 7.1 to 7.9, and about 7.4.

In a specific embodiment, the composition of the subject invention doesnot comprise glucose, glutamine, methionine, and/or lactose.

In one specific embodiment, the composition comprises lysine, glycine,threonine, valine, and tyrosine in a form of free amino acids. In afurther specific embodiment, the composition comprises lysine, glycine,threonine, valine, tyrosine, aspartic acid, isoleucine, tryptophan,asparagine, and serine in a form of free amino acids.

In a further embodiment, the composition comprises one or moredipeptides that are made of the same or different amino acids selectedfrom lysine, glycine, threonine, valine, tyrosine, aspartic acid,isoleucine, tryptophan, asparagine, or serine.

In one embodiment, the composition does not contain glutamine and/ormethionine; and any di-, oligo-, or polypeptides or proteins that can behydrolyzed into glutamine and/or methionine.

In an alternative embodiment, the composition may comprise free aminoacid glutamine, and, optionally, one or more glutamine-containingdipeptides, wherein the total concentration of the free amino acidglutamine and the glutamine-containing dipeptide(s) is less than 300mg/l, or any concentrations lower than 300 mg/l, such as 100 mg/l, 50mg/l, 10 mg/l, 5 mg/l, 1 mg/l, 0.5 mg/l, or 0.01 mg/l.

In another alternative embodiment, the therapeutic composition maycomprise free amino acid methionine, and, optionally, one or moremethionine-containing dipeptides, wherein the total concentration of thefree amino acid methionine and the methionine-containing dipeptide(s) isless than 300 mg/l, or any concentrations lower than 300 mg/l, such as100 mg/l, 50 mg/l, 10 mg/l, 5 mg/l, 1 mg/l, 0.5 mg/l, or 0.01 mg/l.

In one embodiment, the therapeutic composition does not contain anysaccharides, including any mono-, di-, oligo-, polysaccharides, andcarbohydrates. In one specific embodiment, the therapeutic compositiondoes not contain glucose, and/or any di-, oligo, polysaccharides, andcarbohydrates that can be hydrolyzed into glucose. In a specificembodiment, the composition does not contain lactose. In anotherspecific embodiment, the therapeutic composition does not containfructose and/or galactose, and/or any di-, oligo-, polysaccharides, andcarbohydrates that can be hydrolyzed into fructose and/or galactose.

In an alternative embodiment, the therapeutic composition may comprisemonosaccharide glucose, and, optionally, one or more glucose-containingdisaccharides other than lactose, wherein the total concentration of themonosaccharide glucose and the glucose-containing disaccharide(s) isless than 3 g/l, or any concentrations lower than 3 g/l, such as 1 g/l,500 mg/l, 300 mg/l, 100 mg/l, 50 mg/l, 10 mg/l, 5 mg/l, 1 mg/l, 0.5mg/l, or 0.01 mg/l.

In certain embodiments, the therapeutic composition comprises one ormore electrolytes selected from, for example, Na⁺; K⁺; HCO₃ ⁻; CO₃ ²⁻;Ca²⁺; Mg²⁺; Fe²; Cl⁻; phosphate ions, such as H₂PO₄ ⁻, HPO₄ ²⁻, and PO₄³⁻; zinc; iodine; copper; iron; selenium; chromium; and molybdenum. Inan alternative embodiment, the composition does not contain HCO₃ ⁻ orCO₃ ²⁻. In another alternative embodiment, the composition comprisesHCO₃ ⁻ and CO₃ ²⁻ at a total concentration of less than 5 mg/l, orconcentrations lower than 5 mg/l.

In a further embodiment, the therapeutic composition comprises one ormore vitamins including, but not limited to, vitamin A, vitamin C,vitamin D (e.g., vitamin D₁, D₂, D₃, D₄, and/or D₅), vitamin E, vitaminB₁ (thiamine), vitamin B₂ (e.g., riboflavin), vitamin B₃ (e.g., niacinor niacinamide), vitamin B₅ (pantothenic acid), vitamin B₆ (pyridoxine),vitamin B₇ (biotin), vitamin B₉ (e.g., folate or folic acid), vitaminB₁₂ (cobalamin), and vitamin K (e.g., vitamin K₁, K₂, K₃, K₄, and K₅),and choline.

In certain embodiments, the composition does not contain one or more ofthe ingredients selected from oligo-, polysaccharides, andcarbohydrates; oligo-, or polypeptides or proteins; lipids; small-,medium-, and/or long-chain fatty acids; and/or food containing one ormore above-mentioned nutrients.

In one embodiment, phosphate ions, such as H₂PO₄ ⁻, HPO₄ ²⁻, and PO₄ ³⁻,are used to buffer the composition of the subject invention. In oneembodiment, the therapeutic composition uses HCO₃ ⁻ or CO₃ ²⁻ as abuffer. In another embodiment, the therapeutic composition does not useHCO₃ ⁻ or CO₃ ²⁻ as buffer.

The term “consisting essentially of,” as used herein, limits the scopeof the ingredients and steps to the specified materials or steps andthose that do not materially affect the basic and novelcharacteristic(s) of the present invention, i.e., compositions andmethods for improving pulmonary function. For instance, by using“consisting essentially of,” the therapeutic composition does notcontain any unspecified ingredients including, but not limited to, freeamino acids, di-, oligo-, or polypeptides or proteins; and mono-, di-,oligo-, polysaccharides, and carbohydrates that have a direct beneficialor adverse therapeutic effect on pulmonary function. Also, by using theterm “consisting essentially of,” the composition may comprisesubstances that do not have therapeutic effects on pulmonary function;such ingredients include carriers, excipients, adjuvants, flavoringagents, etc that do not affect pulmonary function.

The term “oligopeptide,” as used herein, refers to a peptide consistingof three to twenty amino acids. The term “oligosaccharides,” as usedherein, refers to a saccharide consisting of three to twentymonosaccharides.

Improvement of Pulmonary Function

In one embodiment, the subject invention provides a method for improvingpulmonary function, wherein the method comprises administering, to asubject in need of such treatment, a therapeutic composition of theinvention.

In one embodiment, the composition is formulated for oraladministration. In another embodiment, the composition is formulated forpulmonary administration.

In one specific embodiment, the subject invention provides a method forimproving pulmonary function, and/or for the prevention and/or treatmentof radiation-induced lung injury and lung complications, wherein themethod comprises administering, to a patient or subject in need of suchtreatment, an effective amount of a composition comprising, consistingessentially of, or consisting of one or more free amino acids selectedfrom lysine, glycine, threonine, valine, tyrosine, aspartic acid,isoleucine, tryptophan, asparagine, and serine; one or moreelectrolytes; and optionally, therapeutically acceptable carriers,electrolytes, buffering agents, and flavoring agents.

In one specific embodiment, the subject invention provides a method forimproving pulmonary function, and/or for the prevention and/or treatmentof radiation-induced lung injury and lung complications, wherein themethod comprises administering, to a patient or subject in need of suchtreatment, an effective amount of a composition comprising, consistingessentially of, or consisting of L-lysine, L-glycine, L-threonine,L-valine, L-tyrosine, L-aspartic acid, L-isoleucine, and L-serine; oneor more electrolytes selected from Na⁺, Ca²⁺, Mg²⁺, HCO₃ ⁻, and Cl⁻; andoptionally, therapeutically acceptable carriers, buffering agents, andflavoring agents.

In one embodiment, the subject invention can be used to prevent andtreat lung complications induced by radiation. In certain embodiments,the subject invention can be used to prevent and treat radiation-inducedlung complications including, but not limited to, alveolitis,pneumonitis, and pulmonary fibrosis.

In one embodiment, the therapeutic composition is administered to asubject that received radiation, and the composition can be administeredbefore, during, or after irradiation.

In one embodiment, the subject invention can be used to prevent andtreat lung complications induced by lung injury or lung inflammation. Inone embodiment, the composition can be administered to a patient orsubject immediately before, during, and/or after injury to the lungs,and can be administered once or multiple times each day.

In certain embodiments, the subject invention can be used to prevent andtreat pneumonitis, pulmonary fibrosis and/or other lung diseases orcomplications induced by radiation (such as ionizing radiation),cytotoxic chemotherapeutic agents, proton therapy; pollutants, toxins,trauma, cigarette smoking, autoimmune diseases such as rheumatoidarthritis, medications (e.g., amiodarone, bleomycin, busulfan,methotrexate, and nitrofurantoin), asbestos, and/or infection (e.g.viral, bacterial, fungal and parasitic infection).

In certain embodiments, the present invention can be used to treat lungdiseases including, bronchial asthma, pneumonia, bronchiectasis,interstitial lung diseases, acute and/or chronic pneumonitis, chronicobstructive pulmonary disease (COPD), asthma, silicosis, and lunginjury.

In certain embodiments, the subject invention can be used to preventand/or treat pneumonitis and/or pulmonary fibrosis in subjects thatreceive radiation therapy for cancer or tumor.

In certain embodiments, the subject invention can be used to preventand/or treat pneumonitis and/or pulmonary fibrosis in subjects that areaccidentally exposed to radiation, such as for example, astronauts andpilots who are routinely exposed to space radiation, and subjectsexposed to radiation due to nuclear accident, acts of war, or terrorism.

The term “pulmonary fibrosis” or “lung fibrosis”, as used herein, refersto abnormal formation or accumulation of fibrous, connective, or scartissues and/or matrix macromolecules (e.g., collagens, fibronectins,proteoglycans) on and/or within lungs.

The term “pneumonitis,” as used herein, refers to its ordinary meaning,which is inflammation of lung tissue.

The term “treatment” or any grammatical variation thereof (e.g., treat,treating, and treatment etc.), as used herein, includes but is notlimited to, alleviating a symptom of a disease or condition; and/orreducing, suppressing, inhibiting, lessening, or affecting theprogression, severity, and/or scope of a disease or condition.

The term “prevention” or any grammatical variation thereof (e.g.,prevent, preventing, and prevention etc.), as used herein, includes butis not limited to, delaying the onset of symptoms, preventing relapse toa disease, increasing latency between symptomatic episodes, or acombination thereof. Prevention, as used herein, does not require thecomplete absence of symptoms.

The term “effective amount,” as used herein, refers to an amount that iscapable of treating or ameliorating a disease or condition or otherwisecapable of producing an intended therapeutic effect.

The term “subject” or “patient,” as used herein, describes an organism,including mammals such as primates, to which treatment with thecompositions according to the present invention can be provided.Mammalian species that can benefit from the disclosed methods oftreatment include, but are not limited to, apes, chimpanzees,orangutans, humans, monkeys; domesticated and laboratory animals such asdogs, cats, horses, cattle, pigs, sheep, goats, chickens, mice, rats,guinea pigs, and hamsters.

In one embodiment, the subject or patient in need of treatment of theinvention has received, or will receive, radiation (such as ionizingradiation) at a dose capable of causing lung injury. In certainembodiments, the subject or patient in need of treatment of theinvention has received, or will receive, radiation at a dose of at least1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60,65, 70, 80, 85, 90, 95, or 100 Gy. In certain embodiments, the subjector patient in need of treatment of the invention has received, or willreceive, radiation at a dose at least 0.1, 0.3, 0.5, 0.7, 0.9, 1.0, 1.1,1.2, 1.3, 1.4, 1.5, 1,6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5,2,7, 3.0, 3.2, 3.5, or 4.0 Gy per day.

In one embodiment, the subject or patient in need of treatment of theinvention has received, or will receive, thoracic radiation.

In one embodiment, the subject or patient in need of treatment of theinvention has lung injury, lung inflammation, or lung infection.

In one embodiment, the subject invention does not encompass theprevention or treatment of diseases, disorders, or complications in thegastrointestinal tract. In one embodiment, the subject invention doesnot encompass the prevention or treatment of diseases, disorders, orcomplications disclosed in PCT/US2011/053265, entitled Materials andMethods for Improving Gastrointestinal Function.

Formulations and Administration

The subject invention provides for therapeutic or pharmaceuticalcompositions comprising a therapeutically effective amount of thesubject composition and, optionally, a pharmaceutically acceptablecarrier. Such pharmaceutical carriers can be sterile liquids, such aswater. The therapeutic composition can also comprise excipients,adjuvants, flavoring agents, etc. In an embodiment, the therapeuticcomposition and all ingredients contained therein are sterile.

In one embodiment, the therapeutic composition of the subject inventionis formulated for oral administration. In another embodiment, thetherapeutic composition of the subject invention is formulated forpulmonary administration.

The term “carrier” refers to a diluent, adjuvant, excipient, or vehiclewith which the compound is administered. Examples of suitablepharmaceutical carriers are described in “Remington's PharmaceuticalSciences” by E. W. Martin. Such compositions contain a therapeuticallyeffective amount of the therapeutic composition, together with asuitable amount of carrier so as to provide the form for properadministration to the patient.

The invention also provides a pharmaceutical pack or kit comprising oneor more containers filled with one or more of the ingredients, e.g.,compound, carrier, or the pharmaceutical compositions of the invention.

In certain embodiments, the compositions are prepared in a form adaptedfor delivery into the lungs. For instance, the liquid pharmaceuticalcomposition may be lyophilized prior to use in pulmonary delivery, wherethe lyophilized composition is milled to obtain the finely divided drypowder consisting of particles within a desired size range. For anotherinstance, spray-drying may be used to obtain a dry powder form of theliquid pharmaceutical composition, and the process is carried out underconditions that result in a substantially amorphous finely divided drypowder consisting of particles within the desired size range. Formethods of preparing dry powder forms of pharmaceutical compositions,see, for example, WO 96/32149; WO 97/41833; WO 98/29096; and U.S. Pat.Nos. 5,976,574; 5,985,248; 6,001,336; and 6,875,749 herein incorporatedby reference. In addition, the dry powder form of the pharmaceuticalcomposition may be prepared and dispensed as an aqueous or nonaqueoussolution or suspension, in a metered-dose inhaler.

A surfactant may be added to the pharmaceutical composition to reduceadhesion of the dry powder to the walls of the delivery device fromwhich the aerosol is dispensed. Suitable surfactants for this intendeduse include, but are not limited to, sorbitan trioleate, soya lecithin,and oleic acid. Devices suitable for pulmonary delivery of a dry powderform of a composition as a nonaqueous suspension are commerciallyavailable. Examples of such devices include the Ventolin metered-doseinhaler (Glaxo Inc., Research Triangle Park, N.C.) and the Intal Inhaler(Fisons, Corp., Bedford, Mass.). See also the aerosol delivery devicesdescribed in U.S. Pat. Nos. 5,522,378; 5,775,320; 5,934,272; and5,960,792 herein incorporated by reference.

In yet another embodiment, the pharmaceutical composition can bedelivered in a controlled release system. In one embodiment, a pump maybe used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng.14:201; Buchwald et al., 1980, Surgery 88:507; and Saudek et al., 1989,N. Engl. J. Med. 321:574). In another embodiment, polymeric materialscan be used (see Medical Applications of Controlled Release, Langer andWise (eds.), CRC Pres., Boca Raton, Fla. (1974); Controlled DrugBioavailability, Drug Product Design and Performance, Smolen and Ball(eds.), Wiley, New York (1984); Ranger and Peppas, J. Macromol. Sci.Rev. Macromol. Chem. 23:61 (1983); see also Levy et al., 1985, Science228:190; During et al., 1989, Ann. Neurol. 25:351; Howard et al., 1989,J. Neurosurg. 71:105).

In one embodiment, the pharmaceutical pack or kit further comprisesinstructions for administration, for example, with respect to effectivetherapeutic doses, and/or the timing of administration with referenceto, for example, the elapse time from the exposure to radiation,chemotherapy, or proton therapy. For instance, with regard to subjectsthat receive, or are about to receive, radiation, the therapeutic doseof the composition is determined based on radiation sources, the bodypart being irradiated, and/or the time that has elapsed afterirradiation. With regard to subjects that receive, or are about toreceive chemotherapy, the therapeutic dose of the composition isdetermined based on the type of chemotherapeutic agents, the dosage ofchemotherapeutic agent, and/or the time that has elapsed afterchemotherapy. With regard to subjects that receive, or are about toreceive, proton therapy, the therapeutic dose of the composition isdetermined based on the dosages of proton therapy received by thesubject, and/or the time that has elapsed after proton therapy.

The compositions of the subject invention can be administered to thesubject being treated by standard routes, including oral, inhalation, orparenteral administration including intravenous, subcutaneous, topical,transdermal, intradermal, transmucosal, intraperitoneal, intramuscular,intracapsular, intraorbital, intracardiac, transtracheal, subcutaneous,subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal,epidural and intrasternal injection, infusion, and electroporation, aswell as co-administration as a component of any medical device or objectto be inserted (temporarily or permanently) into a subject.

EXAMPLE

Following is an example that illustrates procedures and embodiments forpracticing the invention. The example should not be construed aslimiting.

Example 1 Therapeutic Compositions for Improving Pulmonary Function

This Example provides formulations for improving lung function, as wellas for prevention and/or treatment of lung complications induced byradiation.

Formulation 1 (Serving Size 1 bottle (237 ml) Amount per serving % DailyValue Calories 10 Chloride 550 mg 16% Sodium 370 mg 15% Salt Blend 929mg Sodium Chloride, Calcium Chloride, Magnesium Chloride L-Valine 276 mgL-Aspartic Acid 252 mg L-Serine 248 mg L-Isoleucine 248 mg L-Threonine225 mg L-Lysine HCL 172 mg L-Glycine 141 mg L-Tyrosine  51 mg OtherIngredients: Water, Natural Flavor, Sodium Bicarbonate

Formulation 2 (Serving Size 1 bottle (237 ml) Amount per servingCalories 10 % Daily Value Total Fat 0 g  0% Sodium 440 mg 18% TotalCarbohydrate 0 g  0% Protein 2 g Ingredients: Water, Amino Acids(L-Tryptophan, L-Valine, L-Aspartic Acid, L-Serine, L-Isoleucine,L-Threonine, L-Lysine Hydrochloride, L-Glycine, L-Tyrosine), Salt,Natural Flavor, Sodium Bicarbonate, Calcium Chloride, Magnesium ChlorideAmount mg/1 bottle Amino Acid serving (237 ml) L-Lysine HCI 175L-Aspartic Acid 255 L-Glycine 144 L-Isoleucine 251 L-Threonine 228L-Tyrosine  52 L-Valine 281 L-Tryptophan 392 L-Serine 252

Mice received radiation at a dose of 8 Gy. Twenty-four hours afterirradiation, mice are treated with the therapeutic composition of thesubject invention for a period of 14 days. Six months after irradiation,pulmonary function test, electrophysiology, radiological andhistopathological examinations are performed.

As shown in FIG. 1A-G, mice treated with the composition of the subjectinvention exhibit improved pulmonary function, electrophysiology,radiological and histopathological features.

All references, including publications, patent applications and patents,cited herein are hereby incorporated by reference to the same extent asif each reference was individually and specifically indicated to beincorporated by reference and was set forth in its entirety herein.

The terms “a” and “an” and “the” and similar referents as used in thecontext of describing the invention are to be construed to cover boththe singular and the plural, unless otherwise indicated herein orclearly contradicted by context.

Recitation of ranges of values herein are merely intended to serve as ashorthand method of referring individually to each separate valuefalling within the range, unless otherwise indicated herein, and eachseparate value is incorporated into the specification as if it wereindividually recited herein. Unless otherwise stated, all exact valuesprovided herein are representative of corresponding approximate values(e.g., all exact exemplary values provided with respect to a particularfactor or measurement can be considered to also provide a correspondingapproximate measurement, modified by “about,” where appropriate).

The use of any and all examples, or exemplary language (e.g., “such as”)provided herein, is intended merely to better illuminate the inventionand does not pose a limitation on the scope of the invention unlessotherwise indicated. No language in the specification should beconstrued as indicating any element is essential to the practice of theinvention unless as much is explicitly stated.

The description herein of any aspect or embodiment of the inventionusing terms such as “comprising”, “having”, “including” or “containing”with reference to an element or elements is intended to provide supportfor a similar aspect or embodiment of the invention that “consists of”,“consists essentially of”, or “substantially comprises” that particularelement or elements, unless otherwise stated or clearly contradicted bycontext (e.g., a composition described herein as comprising a particularelement should be understood as also describing a composition consistingof that element, unless otherwise stated or clearly contradicted bycontext).

It should be understood that the examples and embodiments describedherein are for illustrative purposes only and that various modificationsor changes in light thereof will be suggested to persons skilled in theart and are to be included within the spirit and purview of thisapplication.

We claim:
 1. A method for treating at least one lung complicationinduced by radiation in a human subject in need thereof, wherein themethod comprises: administering, to the human subject in need thereof,an effective amount of a therapeutic formulation comprising free aminoacids, the free amino acids consisting essentially of free amino acidsof lysine, glycine, threonine, valine, tyrosine, aspartic acid,isoleucine, and serine; and optionally, tryptophan; wherein the humansubject in need thereof receives radiation at a dose capable of causinglung injury, wherein the therapeutic formulation is administered viainhalation, and wherein the effective amount is sufficient to improvepulmonary function impaired by the at least one lung complicationinduced by radiation in the human subject in need thereof.
 2. The methodof claim 1, wherein the human subject in need thereof receives radiationin a chest area at a dose capable of causing lung injury.
 3. The methodof claim 1, wherein the therapeutic formulation further comprisestherapeutically acceptable carriers, electrolytes, buffering agents,vitamins, or flavoring agents, or any combination thereof.
 4. The methodof claim 1, wherein the therapeutic formulation consists essentially ofL-lysine, L-glycine, L-threonine, L-valine, L-tyrosine, L-aspartic acid,L-isoleucine, and L-serine; and optionally, therapeutically acceptablecarriers, electrolytes, buffering agents, vitamins, or flavoring agents,or any combination thereof.
 5. The method of claim 1, wherein thetherapeutic formulation comprises one or more electrolytes selected fromNa+, Ca2+, Mg2+, HCO3−, and Cl−.
 6. The method of claim 1, wherein thetherapeutic formulation has a total osmolarity from 165 mOsm to 300mOsm.
 7. The method of claim 1, wherein the therapeutic formulation isadministered before, during, or after irradiation, or any combinationthereof.
 8. The method of claim 1, wherein the lung complication inducedby radiation is radiation-induced alveolitis, radiation-inducedpneumonitis, or radiation-induced pulmonary fibrosis, or any combinationthereof.
 9. The method of claim 1, wherein the human subject in needthereof is a pilot or astronaut.
 10. The method of claim 1, wherein thetherapeutic formulation consists essentially of L-lysine, L-glycine,L-threonine, L-valine, L-tyrosine, L-aspartic acid, L-isoleucine,L-serine, and L-tryptophan; and optionally, therapeutically acceptablecarriers, electrolytes, buffering agents, vitamins, or flavoring agents,or any combination thereof.
 11. The method of claim 10, wherein thetherapeutic formulation comprises one or more electrolytes selected fromNa+, Ca2+, Mg2+, HCO3−, and Cl−.
 12. The method of claim 10, wherein thetherapeutic formulation has a total osmolarity from 165 mOsm to 300mOsm.
 13. The method of claim 10, wherein the therapeutic formulation isadministered before, during, or after irradiation, or any combinationthereof.
 14. The method of claim 10, wherein the lung complicationinduced by radiation is radiation-induced alveolitis, radiation-inducedpneumonitis, or radiation-induced pulmonary fibrosis, or any combinationthereof.
 15. The method of claim 10, wherein the human subject in needthereof is a pilot or astronaut.